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MycoLight™ Green JJ99 *5 mM in DMSO*
MycoLight™ Green JJ99 stain is an excellent green-fluorescent nuclear and chromosome counterstain that is permeant to both prokaryotic and eukaryotic cell membranes. MycoLight™ Green JJ99 stain has a high affinity for DNA and exhibits enhanced fluorescence upon binding with an excitation maximum close to the 488 nm argon laser line and fluorescence emission maximum at ∼500 nm. MycoLight™ Green JJ99 stain is particularly useful as a nuclear counterstain for bacterial assays since it stains both live and dead Gram-positive and Gram-negative bacteria. It is an excellent replacement for SYTO® 9 (SYTO® is the trademark of Invitrogen).
Example protocol
SAMPLE EXPERIMENTAL PROTOCOL
The following protocol can be adapted for most cell types. These conditions require adjustment for each cell type and experimental system. Growth medium, cell density, the presence of other cell types and factors may influence staining. Residual detergent on glassware may also affect staining of many organisms, and cause brightly stained material to appear in solutions with or without cells present.
Use plastic tubes when diluting MycoLight™ Green JJ99, because the diluted stain adheres to glass. In general, the best results are obtained in buffers that do not contain phosphate.
Table 1.Suggested conditions for staining cells with MycoLight™ Green JJ99
Use plastic tubes when diluting MycoLight™ Green JJ99, because the diluted stain adheres to glass. In general, the best results are obtained in buffers that do not contain phosphate.
Table 1.Suggested conditions for staining cells with MycoLight™ Green JJ99
| Application | Concentration | Staining Conditions |
| Bacterial cells | 50 nM – 20 μM | Vortex to mix, then incubate for 1–30 minutes. |
| Eukaryotic cells | 10 nM – 5 μM | Incubate for 10–120 minutes. |
| Microarrays | 50 nM in TE buffer | Incubate for 5 minutes, rinse and then dry. |
- Adherent cells in culture may be stained in situ on coverslips. Pellet cells in suspension by centrifugation and resuspend in buffered salt solution or water.
- Dilute the MycoLight™ Green JJ99 with non-phosphate buffer such as Hepes buffer or buffer of your choice. Add MycoLight™ Green JJ99 using the concentrations listed in Table 1 as a guideline.
Note In initial experiments, it may be best to try several dye concentrations over the entire suggested range to determine the concentration that yields optimal staining. - Stained eukaryotic cells generally show diffuse cytoplasmic staining as well as nuclear staining. Particularly MycoLight™ Green JJ99 show intense staining of intranuclear bodies frequently.
Spectrum
Product family
| Name | Excitation (nm) | Emission (nm) |
| MycoLight™ Green JJ98 *5 mM in DMSO* | 482 | 512 |
| MycoLight™ Green JJ98 | 482 | 512 |
References
View all 25 references: Citation Explorer
A loop-mediated isothermal amplification (LAMP) assay for Strongyloides stercoralis in stool that uses a visual detection method with SYTO-82 fluorescent dye
Authors: Watts MR, James G, Sultana Y, Ginn AN, Outhred AC, Kong F, Verweij JJ, Iredell JR, Chen SC, Lee R.
Journal: Am J Trop Med Hyg (2014): 306
Authors: Watts MR, James G, Sultana Y, Ginn AN, Outhred AC, Kong F, Verweij JJ, Iredell JR, Chen SC, Lee R.
Journal: Am J Trop Med Hyg (2014): 306
A new triplex real time PCR which distinguishes between MRSA, MSSA, and mecA coagulase negative strains by means of melting point analysis using SYTO 9
Authors: Weidner J, Cassens U, Gohde W, Wullenweber J, Greve B.
Journal: Clin Lab (2013): 795
Authors: Weidner J, Cassens U, Gohde W, Wullenweber J, Greve B.
Journal: Clin Lab (2013): 795
Compatibility of SYTO 13 and Hoechst 33342 for longitudinal imaging of neuron viability and cell death
Authors: Hubbard KS, Gut IM, Scheeler SM, Lyman ME, McNutt PM.
Journal: BMC Res Notes (2012): 437
Authors: Hubbard KS, Gut IM, Scheeler SM, Lyman ME, McNutt PM.
Journal: BMC Res Notes (2012): 437
Evaluation of Pseudomonas aeruginosa (PAO1) adhesion to human alveolar epithelial cells A549 using SYTO 9 dye
Authors: Larrosa M, Truchado P, Espin JC, Tomas-Barberan FA, Allende A, Garcia-Conesa MT.
Journal: Mol Cell Probes (2012): 121
Authors: Larrosa M, Truchado P, Espin JC, Tomas-Barberan FA, Allende A, Garcia-Conesa MT.
Journal: Mol Cell Probes (2012): 121
Rapid quantification of cell viability and apoptosis in B-cell lymphoma cultures using cyanine SYTO probes
Authors: Wlodkowic D, Skommer J, Darzynkiewicz Z.
Journal: Methods Mol Biol (2011): 81
Authors: Wlodkowic D, Skommer J, Darzynkiewicz Z.
Journal: Methods Mol Biol (2011): 81
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